Sisk W P, Davis G L, Kingsley D, Chiu A T, Reilly T M
E.I. DuPont de Nemours and Co., Central Research and Development Department, Wilmington, DE 19880.
Gene. 1990 Dec 15;96(2):305-9. doi: 10.1016/0378-1119(90)90269-w.
Segments of a cDNA encoding human plasminogen activator inhibitor type 1 (PAI-1) were subcloned into a highly regulated and inducible Escherichia coli expression system. A plasmid encoding the mature form of human endothelial PAI-1 produced a functional recombinant molecule, as indicated by its ability to inhibit tissue plasminogen activator's enzymatic activity. In contrast to PAI-1 isolated from human fibrosarcoma cells, the biological activity of the recombinant PAI-1 was not dependent on pretreatment with denaturing agents. A construct encoding a polypeptide lacking the first 80 amino acids of PAI-1 also produced elevated levels of the truncated recombinant protein. However, this truncated product was functionally inactive, indicating that an intact N terminus is required for activity.
将编码人1型纤溶酶原激活物抑制剂（PAI-1）的cDNA片段亚克隆到一个高度调控且可诱导的大肠杆菌表达系统中。编码人内皮PAI-1成熟形式的质粒产生了一种功能性重组分子，这可通过其抑制组织纤溶酶原激活物酶活性的能力得以表明。与从人纤维肉瘤细胞中分离出的PAI-1不同，重组PAI-1的生物活性不依赖于用变性剂进行预处理。编码缺失PAI-1前80个氨基酸的多肽的构建体也产生了水平升高的截短重组蛋白。然而，这种截短产物在功能上无活性，表明完整的N末端是活性所必需的。
